1. Field of the Invention
The present invention relates to methods and products useful in restricting the growth, spread and survivability of viruses and bacteria in animals, especially humans. More particularly, the invention relates to methods and medicinal mushroom mycelium products for treating Herpes, Orthopox, influenza, SARS, Hepatitis, Tuberculosis, Escherichia coli and Staphylococcus aureus and other viruses and bacteria.
2. Description of the Related Art
Despite advances in modern medicine, microbes and viruses continue to kill millions of people, stimulating the search for new antimicrobial and antiviral agents, some of which have proven to be of significant commercial value. A major difficulty in the discovery of antimicrobial and antiviral agents is their inherent toxicity to the affected host organism. For instance, a novel agent or treatment that kills the virus but also harms the human host is neither medically practicable nor commercially attractive. Hence, many new antiviral drugs have never made it past preliminary screening studies as they have failed to prove non-toxicity and are unsafe to consume.
That medicinal mushrooms have been ingested for hundreds, and in some cases, thousands of years, is strong support for their non-toxicity, making them appealing candidates in the search for new antimicrobial and antiviral agents. The cell surface of mycelium secretes antibiotics in a kind of “sweat” which are known in the field as exudates or secondary metabolites. These antibiotics and enzymes target distinct sets of microbes. Useful antibiotics isolated from mushrooms include calvacin from the Giant Puffball (Calvatia gigantea), armilliaric acid from Honey Mushrooms (Armillaria mellea), campestrin from Agaricus campestris (The Meadow Mushroom), coprinol from Inky Caps (Coprinus species) corolin from Turkey Tail Mushrooms (Trametes versicolor=Coriolus versicolor), cortinellin from Shiitake (Lentinula edodes), ganomycin from Reishi (Ganoderma lucidum) and sparassol from Cauliflower mushrooms (Sparassis crispa).
Suzuki et al. (1990) characterized an antiviral water-soluble lignin in an extract of the mycelium of Shiitake mushrooms (Lentinula edodes) isolated from cultures grown on rice bran and sugar cane bagasse which limited HIV replication in vitro and stimulated the proliferation of bone-marrow cells. Clinical trials with lentinan in the treatment of HIV patients showed inhibitory activity. (Gordon et al., 1998). However, Abrams (2002) found no significant advantage in using lentinan in treating AIDS patients. Another mushroom recognized for its antiviral activity is Fomes fomentarius, a hoof-shaped wood conk growing trees, which inhibited the tobacco mosaic virus (Aoki et al., 1993). Collins & Ng (1997) identified a polysaccharopeptide inhibiting HIV type 1 infection from Turkey Tail (Trametes versicolor) mushrooms while Sarkar et al. (1993) identified an antiviral substance resident in an extract of Shiitake (Lentinula edodes) mushrooms. More recently, derivatives of the Gypsy mushroom, Rozites caperata, were found by Piraino & Brandt (1999) to have significant inhibition against the replication and spread of varicella zoster (the ‘shingles’ and ‘chickenpox’ virus), influenza A, and the respiratory syncytial virus (RSV) but not against HIV and other viruses. Eo et al. (1999) found antiviral activity from the methanol-soluble fractions of Reishi mushrooms (Ganoderma lucidum), selectively inhibiting Herpes simplex and the vesicular stomatitus virus (VSV). Wang & Ng (2000) isolated a novel ubiquitin-like glycoprotein from Oyster mushrooms (Pleurotus ostreatus) that demonstrated inhibitory activity toward the HIV-1 reverse transcriptase. Arabinoxylane inhibits HIV indirectly through the enhancement of NK cells that target the virus. Arabinoxylanes are created from mushroom mycelia's enzymatic conversion of rice bran (Ghoneum, M., 1998). Research by Dr. Byong Kak Kim showed that extracts of Reishi (Ganodenna lucidum) prevented the death of lymphocytes infected with HIV and inhibited the replication of the virus within the mother and daughter cells (Kim et al., 1994). In response to hot water extracts of Reishi mushrooms, preserved in ethanol, versus saline controls, NK cell activity was significantly augmented when cancer cells were co-cultured with human spleen cells. (Ohmoto, 2002). A mycelial combination of 7 species grown on rice achieved a similar result, greater than any one species at the same dosage. As the water extract of the fruitbodies is high in beta glucans while the mycelium-on-rice is low in beta glucans, but is high in arabinoxylanes, two causal agents are identified as NK effectors. Both the extract and the heat treated, freeze dried, powdered mycelium from 7 species share common activity levels of enhancing NK activity by 300+%. These compounds may be synergistic. This same combination of 7 species fermented on rice had a strong effect against HIV, inhibiting replication by 99% while the water extract of Reishi fruitbodies was 70%, respectively. These results underscore that water extractions of fruitbodies and oral administration of myceliated rice positively influence the immune system, activating different subsets of immunological receptor sites. Maitake (Grifola frondosa) is currently the subject of research in the treatment of HIV. Mizuno et al. (1996) noted that crude fractions from Chaga (Inonotus obliquus) showed antiviral activity against HIV.
Fomitopsis officinalis (Villars) Bondarzew & Singer (=Agaricum officinalis, Fomes officinalis, Fomes laricis and Laricifomes officinalis) has the common names Agarikon, Quinine Conk, Larch Bracket Mushroom, Brown Trunk Rot, Eburiko, Adagan (‘ghost bread’) and Tak'a di (‘tree biscuit’). Once widespread throughout the temperate regions of the world, this perennial wood conk saprophytizes larch, Douglas fir and hemlock, preferring mature woodlands. Now nearly extinct in Europe and Asia, this mushroom is a resident of the Old Growth forests of Northern California, Oregon, Washington and British Columbia. Known constituents include beta glucans, triterpenoids and agaricin. Forms used include mushroom fruitbodies and mycelium. F. officinalis has traditionally been used for centuries for the treatment of “coughing illnesses.” Mizuno et al. (1995) and Hanssen (1996) include this mushroom in a group of polypores, the hot water extracts of which provide a strong host mediated response. Agarikon was also applied topically, in a poultice, as an anti-inflammatory and to treat muscle/skeletal pain. Described by the first century Greek physician Dioscorides in Materia Medica, the first encyclopedic pharmacopoeia on the medicinal use of plants, in approximately 65 C.E., as a treatment for a wide range of illnesses, most notably consumption, an archaic medical term. It was not until the invention of the microscope did germ-theory suggest that infections were caused by microbes. A resident on the old growth conifers, especially spruce, hemlock, Douglas fir and on Larch, this amazing mushroom produces a chalky cylindrical fruitbody that adds layers of spore-producing pores with each growth season, allowing for a rough calculation of age. Conks up to 50 years have been collected, and often times they resemble a woman, reminiscent of the Venus of Willendorf form. The Haida First Peoples of the Queen Charlotte Islands, and elsewhere on the coast of British Colombia, associated this mushroom, or debatably another polypore species, with the powerful creator spirit Raven, and as a protector of women's sexuality (Blanchette et al., 1992; Stamets, 2002). This mushroom was carved into animalistic forms and placed on shaman's graves to protect them from evil spirits. Grzywnowicz (2001) described the traditional use of this mushroom by Polish peoples, as a treatment against coughing illnesses, asthma, rheumatoid arthritis, bleeding, infected wounds, and was known for centuries as a “elixirium ad longam vitam”: elixir of long life. The North Coast First Peoples of Northwestern North America also discovered the use of this mushroom as a poultice to relieve swellings and in teas for treating feverish illnesses. Called the Quinine Fungus in many forestry manuals because of its bitter taste, this mushroom is not the source of quinine, an alkaloid from the bark of the Amazonian Cinchona ledgeriana tree which was widely used since the late 19th century to treat malaria, caused by Plasmodium falciparum. Despite the long history of use, few modern studies have been published on its medicinally active compounds. F. officinalis merits further research as the number of strains is in rapid decline, especially in Europe, where it is on the verge of extinction (Leck, 1991).
The present inventor incorrectly speculated that it is thought, but not yet proven, that Fomitopsis officinalis provided an aid in preventing the scourge of viral diseases such as smallpox among native populations of northwestern North America (Stamets 2002). Upon further investigation, the inventor contacted Guujaaw (2004), President of the Haida People who told him “We did not have time to develop a defense against smallpox. Our people went from 50,000 to 500 in three years. The smallpox came from a passenger dropped from the ship, the Queen Charlotte. Had we known of a cure, we would have used it.” Moreover, tests of the hot-water extract from boiling this mushroom showed no antiviral activity with the U.S. Defense Department's Bioshield BioDefense Program whilst the water/ethanol extract from the in vitro grown mycelium originating from a tissue clone of this mushroom showed strong anti-pox virus activity (U.S. patent application Ser. No. 11/029,861).
Piptoporus betulinus (Bull.:Fr.) Karst (=Polyporus betulinus (Bull.:Fr.) Fr.) is commonly known as the Birch Polypore or Kanbatake. It is found throughout the birch forests of the world, circumboreal, and is one of the most common mushrooms on that host. Known constituents include betulin, betulinic acid, agaric acid, single stranded RNA, heteroglucans, and antibiotics. Forms used include mushrooms, mycelium on grain and fermented mycelium. Crude extracts and purified fraction are tumor inhibiting in vitro. The novel antibiotic, Piptamine, has been isolated from this fungus (Schlegel et al. 2000). Pisha et al. (1995) found, in mice studies, that betulinic acid, a pentacyclic triterpene, was specifically toxic to melanoma without adverse effects to the host. Farnsworth et al. (1995) found that betulinic acid facilitated apoptosis of melanoma. This compound has been further evaluated for the treatment or prevention of malignant melanoma. Manez et al. (1997) found that selected triterpenoids reduced chronic dermal inflammation. Found with the famous Ice Man, the use of P. betulinus transcends cultures and millennia. A fungus useful to stop bleeding, prevent bacterial infection, and as an antimicrobial agent against intestinal parasites, this species is one of the most prominent and frequently encountered mushroom seen on birch. Capasso (1998) postulated that the Ice Man used this fungus to treat infection from intestinal parasites (Trichuris trichiura).
Summaries of the antiviral properties of mushrooms were published by Suay et al. (2000), Brandt & Piraino (2000) and Stamets (2001, 2002). Besides having a direct antiviral or antimicrobial effect, mushroom derivatives can also activate natural immune response, potentiating host defense, and in effect have an indirect but significant activity against infections. (Stamets, 2003).
As mushrooms share a more common evolutionary history with animals than with any other kingdom, mushrooms and humans suffer from common pathogens in the microbial world, for instance, the bacterium Staphylococcus aureus and Pseudomonas flourescens. Mushrooms have a vested evolutionary interest in not being rotted by bacteria, producing antibacterial agents to stave off infection. Work by Suay et al. (2000) showed that various mushroom species have anti-bacterially specific properties. Viral infections, as in viral pneumonia, can precede, for instance, bacterial infections from Streptococcus pneumoniae or Staphylococcus aureus, so the use of mushrooms having antibacterial properties can help forestall secondary infections from opportunistic pathogens. Mushrooms having both antibacterial and antiviral properties are especially useful for preventing infection. Furthermore, it is anticipated that some mushrooms will demonstrate anti-bacteriophagic properties, being dually antibacterial and antiviral.
Mushrooms have within them polysaccharides, glycoproteins, ergosterols, enzymes, acids and antibiotics, which individually and in concert can mitigate viral infection. As each species of mushrooms is unique, not only in its cellular architecture, but also in its innate response to viral antagonists, animals, especially humans, can benefit from these antiviral mushroom-derived agents. Since humans now face multiple threats from numerous viruses, including but not limited to HIV, Pox (such as small pox), West Nile virus, influenza and avian or bird flu viruses, coronaviruses such as SARS, hepatitis, Lyme disease, HELA cervical virus, respiratory syncytial virus, hantavirus, vesicular stomatitus, Herpes, Epstein Barr, Varicella-Zoster, Polio, Yellow Fever, Marburg, Ebola, VEE, Lassa and Dengue Fever, and numerous microbes including Plasmodium falciparum, Bacillus anthracis, Escherichia coli, anthrax, Mycobacterium tuberculosis, bacteriophages, fungi such as Candida albicans, Aspergillus, Fusarium, Stachybotrys and Thernoactinomycetes, as well as prions such as BSE, finding substances that afford a broad shield of protection against multiple viruses and microbes is difficult. Virologists are increasingly concerned about the threat of viral infection from animal hosts, thought to be the probable source of the 2003 SARS (Sudden Acute Respiratory Syndrome) epidemic, likely to have originated in rural regions of China where humans and captured animals exist in close quarters. Furthermore, the concentration of animals in ‘factory farms’ wherein thousands of chickens, hogs, cows and other animals are aggregated, provide a breeding environment for contagions as well as other environmental catastrophes. Viruses and bacteria can also breed when birds, dogs, prairie dogs, vermin, cats, primates, bats and other animals, including humans, have concentrated populations. These sources, and more yet to be discovered, present a microbial threat to human health.
Smallpox is a serious acute, contagious and infectious disease marked by fever and a distinctive progressive skin rash. The majority of patients with smallpox recover, but death may occur in up to 30% of cases. Many smallpox survivors have permanent scars over large areas of their body, especially their face, and some are left blind. Occasional outbreaks of smallpox have occurred for thousands of years in India, western Asia and China. European colonization in both the Americas and Africa was associated with extensive epidemics of smallpox among native populations in the 1500s and 1600s, including use as a potential biological weapon in the United States. Smallpox was produced as a weapon by several nations well past the 1972 Bioweapons convention that prohibited such actions.
There is no specific treatment for smallpox and the only prevention is vaccination. In 1980, the disease was declared eradicated following worldwide vaccination programs. However, in the aftermath of the terrorist and anthrax attacks of 2001, the deliberate release of the smallpox virus is now regarded as a possibility and the United States is taking precautions to deal with this possibility.
Smallpox is classified as a Category A agent by the Centers for Disease Control and Prevention. Category A agents are believed to pose the greatest potential threat for adverse public health impact and have a moderate to high potential for large-scale dissemination. Other Category A agents are anthrax, plague, botulism, tularemia, and viral hemorrhagic fevers. Even the remote potential for release of a deadly communicable disease in an essentially non-immune population is truly frightening.
Orthopox (Orthopoxvirus) includes the virus that causes smallpox (Variola major). Smallpox infects only humans in nature, although other primates have been infected in the laboratory. Other members of the Orthopoxvirus genera capable of infecting humans include monkeypox, camelpox, cowpox, and vaccinia. Other poxviruses capable of infecting humans include the Parapoxvirus pseudocowpox and Orf (Parapoxvirus ovis) and the Molluscipoxvirus Molluscum contagiosum. Monkeypox is a rare smallpox-like disease encountered in villages in central and west Africa. It is transmitted by monkeys, primates and rodents. Camelpox is a serious disease of camels. The genetic sequence of the camelpox virus genome is most closely related to that of the Variola (smallpox) virus. Cowpox is usually contracted by milking infected cows and causes ulcerating “milker's nodules” on the hands of dairy workers. Cowpox protects against smallpox and was first used for vaccination against smallpox. Pseudocowpox is primarily a disease of cattle. In humans it causes non-ulcerating “milker's nodes.” Molluscum contagiosum causes minor warty bumps on the skin. It is transferred by direct contact, sometimes as a venereal disease. Orf virus occurs worldwide and is associated with handling sheep and goats afflicted with “scabby mouth.” In humans it causes a single painless lesion on the hand, forearm or face. Vaccinia, a related Orthopox of uncertain origin, has replaced cowpox for vaccination. Other viruses of the Poxyiridae family include buffalopox virus, rabbitpox virus, avipox virus, sheep-pox virus, goatpox virus, lumpy skin disease (Neethling) virus, swinepox virus and Yaba monkey virus.
Poxviruses are very large rectangular viruses the size of small bacteria. They have a complex internal structure with a large double-stranded DNA genome enclosed within a “core” that is flanked by two “lateral bodies.” The surface of the virus particle is covered with filamentous protein components, giving the particles the appearance of a ball of knitting wool. The entire virus particle is encapsulated in an envelope derived from the host cell membranes, thereby “disguising” the virus immunologically. Most poxviruses are host-species specific, but Vaccinia is a remarkable exception. True pox viruses are antigenically rather similar, so that infection by one elicits immune protection against the others.
Influenza (“flu”) is an infection of the respiratory system characterized by fever, body aches, chills, dry cough, headache, sore throat and stuffy nose. The flu, which is caused by a variety of viruses, is notable for its ability to sweep through entire communities in both developed and developing countries and is associated with high morbidity and a significant death rate. Half the population of a community may be affected during an epidemic. Children are much more likely than adults to get sick from the flu, as are families with school-age children—schools are an excellent place for flu viruses to infect and spread. The risk of death from influenza is highest among persons aged 65 or older, although young children, particularly the newborn, and persons with certain chronic conditions are also at risk of death. The flu is particularly serious because of the rapidity of outbreaks, the large number of people affected and the possibility of serious complications such as pneumonia. The Centers for Disease Control and Prevention estimates that 5-20% of the population of the United States come down with the flu each flu season (typically late fall through winter). Although most recover from the illness, according to CDC estimates about 19,000-36,000 died from the flu and its complications each year during the epidemics occurring from 1976-1999. The 1918 Spanish flu pandemic is estimated to have caused 20-40 million deaths worldwide, including 500,00 in the United States. The majority of the 1918 deaths were caused by secondary infections from bacteria, which exploited the scarred lung tissue and immune impairment. The 1957 Asian flu and the 1968 Hong Kong flu outbreaks killed hundreds of thousands in the United States.
The influenza viruses are RNA viruses belonging to the Orthomyxoviridae family. Influenza viruses are classified into types A, B and C. Type A is the most common and usually causes the most serious epidemics. Influenza A viruses are further divided into subtypes on the basis of two proteins found on the surface of the virus, hemagglutinin (H) and neuraminidase (N). Influenza A viruses are found in many different animals, including birds, pigs, whales and seals, with wild birds acting as the reservoir for all subtypes of influenza A viruses. The influenza A subtypes H1N1 and H3N2 have circulated widely among people (the Spanish flu was a H1N1 virus and the Hong Kong flu was a H3N2 virus). Type B can also cause epidemics, but generally produces a milder disease than that caused by type A. Type C viruses have never been connected with major epidemics. Yearly flu vaccines are available targeting new variant strains resulting from antigenic drift, but neither prior vaccination nor previous infection guarantees protection from the flu since the virus typically varies from year to year.
It is currently feared that a strain of avian influenza (“bird flu”), which naturally occurs in wild birds and can spread to domesticated birds, could mutate into a form easily transmissible by human-to-human and cause a worldwide pandemic. The H5N1 high pathogenicity avian influenza (HPAI) virus strain, which is becoming endemic in various Asian countries and has spread to a number of countries in the Middle East, Africa and Europe, has particularly concerned researchers because it is spread by migratory wildfowl, because it is especially virulent and has caused the death of millions of animals worldwide, because it mutates rapidly and continues to evolve and because it has spread to domesticated birds and mammals including pigs and tigers and in limited circumstances to humans. As influenza type A H5 hemagglutinin viruses have not circulated among humans and most or all of the population has no protective antibodies, there is the potential that H5N1 could cause a pandemic were it to mutate to a form easily transmissible by human-to-human contact. The H5N1 avian influenza strain has caused illness in more than several hundred people in Asia and the Middle East, approximately half of whom have died (almost all cases are thought to be the result of bird-to-human infection, but it appears there may be rare cases of human-to-human transmission). A severe influenza pandemic could potentially result in unprecedented death, social disruption and economic loss as millions become seriously ill at the same time.
SARS is a new viral illness spread mainly by close person-to-person contact and possibly by infected surfaces or objects or an airborne vector or other means. SARS is believed to have originated in rural China in November 2002. In March 2003 the alarming spread of cases caused the World Health Organization and U.S. Centers for Disease Control and Prevention to issue a global alert over cases of atypical pneumonia that did not appear to respond to treatment. The illness was named Severe Acute Respiratory Syndrome (SARS). By the third week of March 2003, researchers from several countries had isolated a novel single-stranded RNA virus from the Coronavirus family (SARS-CoV) with contagiousness and high mortality rate unlike any other known human coronaviruses. Although coronaviruses account for about thirty percent of respiratory illnesses, most are moderate in course (such as common colds) with pneumonia being caused only in patients with poor immune systems; SARS-CoV seemed to be the first Coronavirus that consistently caused severe disease in humans. Before the outbreak was contained, it spread to more than two dozen countries. By December of 2003, 774 people had died and more than 8,000 had been infected. World airlines were hit hard by the SARS epidemic as several carriers slashed flights and axed jobs. The tourism industry suffered badly due to the fear unleashed by the outbreak, as did many other businesses and industries far from its epicenter. In many ways SARS caused the worst economic crisis in Southeast Asia since the wave of bank failures and currency devaluations that occurred there in 1988.
SARS causes a form of lung injury characterized by increased permeability of the alveolar-capillary membrane, diffuse alveolar damage, the accumulation of proteinaceous pulmonary edema and pulmonary failure. Symptoms included high fever and one or more respiratory symptoms including, cough, shortness of breath and difficulty breathing. In addition to fever and respiratory symptoms, SARS was associated with other symptoms including headache, muscular stiffness, loss of appetite, malaise, confusion, rash, diarrhea and low oxygen levels in the blood (hypoxia). In many cases, those symptoms were followed by pneumonia in both lungs, sometimes requiring use of a respirator. The pathology of SARS is not yet fully understood and the clinical symptoms are unusual. The disease was mild in children and the mortality rate in that group almost nonexistent. Persons who suffered from chronic disease and the elderly had a much higher mortality rate. Patients who survived SARS infections recovered seemingly spontaneously while those who perished succumbed to rapid respiratory decline accompanied by extensive lung tissue damage. The tissue damage appeared to be driven by the patient's own immune system rather than the organism itself. The mechanism of SARS pathogenesis may involve both direct viral cytocidal effects on the target cells and immune-mediated mechanisms. There are no specific therapies for SARS. The use of physiologically targeted strategies of mechanical ventilation and intensive care unit management including fluid management and glucorticoids was the only supportive therapy available. Numerous antibiotic therapies were tried with no clear effect. Ribavirin with or without use of steroids was used in a number of patients. But, in the absence of clinical indicators, its effectiveness was not proven.
SARS was a much more virulent strain than most coronaviruses, leading scientists to believe that the virus had its origins in a non-human animal, where a coronavirus can have more severe effects. Although this virus most likely originated from a wild animal, perhaps the civet cat, the SARS virus was well adapted in humans as evidenced by the high person-to-person transmissibility of the virus. The critical questions are whether there is extensive horizontal transmission between animals, and whether the jump of the virus from animals to human was a rare and accidental event or portends frequent occurrences in the future. The answers to these questions will determine whether animals are viable reservoirs for future SARS outbreaks and whether person-to-person transmission of SARS-CoV might recur.
With the flow of airline passengers from remote regions of the world, concentrating in airports and being re-routed to their destinations, the contagiousness of foreign-borne viruses carried by passengers are likely to be exacerbated in these types of locations, especially within the closed compartments of passenger airplanes, increasing the likelihood of cross-infection. Virtually anywhere humans concentrate provide opportunities for contagions to spread, whether by air or by physical contact. The history of viruses indicates the danger posed by new strains for which no immunities or vaccines exist. With the increased threat of bioterrorism from weaponized viruses, a readily available broad-spectrum antiviral serves the best interests of public health.